HPLC Analysis of Cortocisteroids in Urine on Discovery^® HS F5 after SPE using Discovery^® DSC-CN

sample preparation

SPE (Solid Phase Extraction)

sample/matrix

human urine spiked with corticosteroids at 0.5 and 1.0 μg/mL and diluted in DI water (1:1, v/v)

SPE well plate

Discovery DSC-CN 96-well SPE, 100 mg/well (575636-U)

condition

1 mL methanol; 1 mL DI water.

sample addition

1 mL spiked urine

washing

1 mL 20% methanol

elution

1 mL methanol

eluate post-treatment

Evaporate eluate with nitrogen at 30 °C. Reconstitute in 200 μL mobile phase.

column

Discovery HS F5, 5 cm x 4.6 mm I.D., 3 μm particles (567504-U)

mobile phase

[A] methanol; [B] DI water; (40:60, A:B)

flow rate

1.5 mL/min

column temp.

35 °C

detector

UV, 240 nm

injection

5 μL